LOCUS       sequence-263384-        4717 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Used as a donor vector to clone into pSLIK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4717)
  AUTHORS   Pereira EJ, Burns JS, Lee CY, Marohl T, Calderon D, Wang L, Atkins 
            KA, Wang CC, Janes KA
  TITLE     Sporadic activation of an oxidative stress-dependent NRF2-p53 
            signaling network in breast epithelial spheroids and 
            premalignancies.
  JOURNAL   Sci Signal. 2020 Apr 14;13(627). pii: 13/627/eaba4200. doi: 
            10.1126/scisignal.aba4200.
  PUBMED    32291314
REFERENCE   2  (bases 1 to 4717)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci 
            Signal."; date: "2020-04-14"; volume: "13(627). pii: 
            13/627/eaba4200. doi"; pages: " 10.1126/scisignal.aba4200"
COMMENT     Sequence Label: pEN_TTmiRc2_BirA
FEATURES             Location/Qualifiers
     source          1..4717
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     25..48
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    67..166
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        198..512
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    242..260
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    277..295
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    313..331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    349..367
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    384..402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    420..438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     485..509
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(519..869,870..872,873..1481)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     protein_bind    complement(1487..1586)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1698..1717)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1776..1795)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2369..2902)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3091..3096,3097..3113,3114..3119))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3776..4364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4265..4284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4529..4615
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc
       61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
      121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat
      181 tcagtcgact ggatcccgag tttactccct atcagtgata gagaacgtat gtcgagttta
      241 ctccctatca gtgatagaga acgatgtcga gtttactccc tatcagtgat agagaacgta
      301 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      361 atagagaacg tatgtcgagt ttatccctat cagtgataga gaacgtatgt cgagtttact
      421 ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt acggtgggag gcctatataa
      481 gcagagctcg tttagtgaac cgtcagatcg ccactagaat gaaggacaac accgtgcccc
      541 tgaagctgat cgccctgctg gccaacggcg agttccactc tggcgagcag ctgggagaga
      601 ccctgggaat gagcagagcc gccatcaaca agcacatcca gacactgaga gactggggag
      661 tggacgtgtt caccgtgcct ggcaagggct acagcctgcc tgagcctatc cagctgctga
      721 acgccaagca gatcctggga cagctggatg gcggaagcgt ggccgtgctg cctgtgatcg
      781 actccaccaa tcagtacctg ctggacagaa tcggagagct gaagtccggc gacgcctgca
      841 tcgccgagta ccagcaggct ggcagaggag gcagaggacg gaagtggttc agcccattcg
      901 gagccaacct gtacctgtcc atgttctgga gactggagca gggacctgct gctgccatcg
      961 gactgagtct ggtgatcgga atcgtgatgg ccgaggtgct gagaaagctg ggagccgaca
     1021 aggtgagagt gaagtggcct aatgacctgt acctccagga ccgcaagctg gctggcatcc
     1081 tggtggagct gacaggcaag acaggcgatg ccgctcagat cgtgatcgga gccggaatca
     1141 acatggccat gagaagagtg gaggagagcg tggtgaacca gggctggatc accctgcagg
     1201 aggctggcat caacctggac cggaacaccc tggccgccat gctgatcaga gagctgagag
     1261 ccgctctgga gctgttcgag caggagggac tggctcctta cctgagcaga tgggagaagc
     1321 tggacaactt catcaacaga cctgtgaagc tgatcatcgg cgacaaggaa atcttcggca
     1381 tctccagagg aatcgacaag cagggagctc tgctgctgga gcaggacgga atcatcaagc
     1441 cctggatggg cggagaaatc tccctgagaa gcgcagagaa gactagaccc agctttcttg
     1501 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1561 atcagtcaaa ataaaatcat tatttgccat ccagctgcag ctctggcccg tgtctcaaaa
     1621 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1681 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     1741 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1801 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     1861 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1921 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1981 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2041 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg tccctgcgcc ggttgcattc
     2101 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2161 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2221 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2281 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaggccg
     2341 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     2401 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     2461 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     2521 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga
     2581 tatagatctc actacgcggc tgctcaaacc tgggcagaac gtaagccgcg agagcgccaa
     2641 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     2701 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     2761 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     2821 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     2881 catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     2941 taacgcgctt gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa
     3001 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     3061 ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac
     3121 tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc
     3181 gaagtcgagg catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat
     3241 cgtcaggcat tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc
     3301 tggcttcagg agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg
     3361 gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac
     3421 tctagctata gttctagtgg ttggctacta ataggttgta ttgatgttgg acgagtcgga
     3481 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     3541 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3601 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     3661 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     3721 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3781 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3841 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3901 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3961 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4021 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4081 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4141 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4201 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4261 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4321 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4381 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4441 cccctgattc tgtggataac cgtattaccg ctagcatgga tctcggggac gtctaactac
     4501 taagcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cggaagactg
     4561 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     4621 gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     4681 ataaactgcc aggcatcaaa ctaagcagaa ggccatc
//
